Relative abundance and ranges of selected diatoms from Pliocene-Pleistocene sections of ODP Leg 177, Atlantic sector of the Southern Ocean

During Ocean Drilling Program (ODP) Leg 177, seven sites were drilled aligned on a transect across the Antarctic Circumpolar Current in the Atlantic sector of the Southern Ocean. The primary scientific objective of Leg 177 was the study of the Cenozoic paleoceanographic and paleoclimatic history of the southern high latitudes and its relationship with the Antarctic cryosphere development. Of special emphasis was the recovery of Pliocene-Pleistocene sections, allowing paleoceanographic studies at millennial or higher time resolution, and the establishment of refined biostratigraphic zonations tied to the geomagnetic polarity record and stable isotope records. At most sites, multiple holes were drilled to ensure complete recovery of the section. A description of the recovered sections and the construction of a multihole splice for the establishment of a continuous composite is presented in the Leg 177 Initial Reports volume for each of the sites (Gersonde, Hodell, Blum, et al., 1999). Here we present the relative abundance pattern and the stratigraphic ranges of diatom taxa encountered from shore-based light microscope studies completed on the Pliocene-Pleistocene sequences from six of the drilled sites (Sites 1089-1094). No shore-based diatom studies have been conducted on the Pliocene-Pleistocene sediments obtained at Site 1088, located on the northern crest of the Agulhas Ridge, because of the scattered occurrence and poor preservation of diatoms in these sections (Shipboard Scientific Party, 1999b). The data included in our report present the baseline of a diatom biostratigraphic study of Zielinski and Gersonde (2002), which (1) includes a refinement of the southern high-latitude Pliocene-Pleistocene diatom zonation, in particular for the middle and late Pleistocene, and (2) presents a biostratigraphic framework for the establishment of age models of the recovered sediment sections. Zielinski and Gersonde (2002) correlated the diatom ranges with the geomagnetic polarity record established shipboard (Sites 1090 and 1092) (Shipboard Scientific Party, 1999c, 1999d) and on shore (Sites 1089, 1091, 1093, and 1094) by Channell and Stoner (2002). The Pliocene-Pleistocene diatom zonation proposed by Zielinski and Gersonde (2002) relies on a diatom zonation from Gersonde and Bárcena (1998) for the northern belt of the Southern Ocean. Because of latitudinal differentiation of sea-surface temperature, nutrients, and salinity between Antarctic and Subantarctic/subtropical water masses, the Pliocene-Pleistocene stratigraphic marker diatoms are not uniformly distributed in the Southern Ocean (Fenner, 1991; Gersonde and Bárcena, 1998). As a consequence, Zielinski and Gersonde (2002) propose two diatom zonations for application in the Antarctic Zone south of the Polar Front (Southern Zonation, Sites 1094 and 1093) and the area encompassing the Polar Front Zone (PFZ) and the Subantarctic Zone (Northern Zonation, Sites 1089-1092). This accounts especially for the Pleistocene zonation where Hemidiscus karstenii, whose first abundant occurrence datum and last occurrence datum defines the subzonation of the northern Thalassiosira lentiginosa Zone, occurs only sporadically in the cold-water realm south of the PFZ and thus is not applicable in sections from this area. However, newly established marker species assigned to the genus Rouxia (Rouxia leventerae and Rouxia constricta) are more related to cold-water environments and allow a refinement of the Pleistocene stratigraphic zonation for the southern cold areas. A study relying on quantitative counts of both Rouxia species confirms the utility of these stratigraphic markers for the identification of sequences attributed to marine isotope Stages 6 and 8 in the southern Southern Ocean (Zielinski et al., 2002).

Chemobiostratigraphy of the Cretaceous/Paleocene boundary at ODP Hole 120-750A

An integrated biostratigraphic and stable isotope investigation was conducted on a high-latitude sequence across the Cretaceous/Paleogene (K/P) boundary recovered in Hole 750A in the southern Indian Ocean. The sequence consists of nannofossil chalk and is discontinuous across the boundary; missing is an estimated 0.3-m.y. late Maestrichtian and early Danian interval. Nonetheless, because calcareous nannofossil Zones NP1 and NP2 are well-developed, micropaleontological studies of the sequence have yielded a detailed record of Danian high-latitude microplankton evolution. In addition, stable carbon isotope analyses of planktonic and benthic foraminifer and bulk samples provide a record of late Maestrichtian and early Danian surface- and deep-water carbon isotope variations. Together, the carbon isotope and carbonate accumulation records serve as an index of regional marine net productivity across the boundary. Earliest Danian nannoplankton assemblages consisted mainly of persistent genera that were generally rare or absent in the Upper Cretaceous at Hole 750A. However, by 0.5-0.6 m.y. after the boundary, newly evolving Danian taxa became dominant. The turnover in nannofossil assemblages was accompanied by significant changes in rates of net productivity as gauged by carbon isotope distributions and carbonate accumulation rates. During the period dominated by persistent taxa, net productivity was extremely low, as reflected by the absence of vertical delta13C gradients and reduced carbonate accumulation rates. Later in the Danian, as new species evolved and flourished, vertical delta13C gradients reappeared and carbonate accumulation rates increased, signaling partial recovery of net productivity in this region. The absolute timing and magnitude of late Maestrichtian and early Danian biotic and geochemical changes in the southern Indian Ocean were similar to those recorded in other pelagic K/P boundary sequences from low- and mid-latitude Atlantic and Pacific sites, indicating that these events were ubiquitous.

Abundance and biomass of phytoplankton in the western part of the Black Sea in September 1991 during the NATO Programme Hydroblack

The samples were concentrated down to 50 cm**3 by slow decantation after storage for 20 days in a cool and dark place. The species identification was done under light microscope OLIMPUS-BS41 connected to a video-interactive image analysis system at magnification of the ocular 10X and objective - 40X. A Sedgwick-Rafter camera (1ml) was used for counting. 400 specimen were counted for each sample, while rare and large species were checked in the whole sample (Manual of phytoplankton, 2005). Species identification was mainly after Carmelo T. (1997) and Fukuyo, Y. (2000). Total phytoplankton abundance was calculated as sum of taxon-specific abundances. Total phytoplankton biomass was calculated as sum of taxon-specific biomasses. The cell biovolume was determined based on morpho-metric measurement of phytoplankton units and the corresponding geometric shapes as described in detail in (Edier, 1979).

Campanian-Maestrichtian planktonic foraminifera of the southern South Atlantic

Campanian-Maestrichtian planktonic foraminifers were examined from Sites 698 (2128 m water depth) and 700 (3611 m water depth) on the Northeast Georgia Rise (southern South Atlantic, 51°S). Site 698 penetrated 72.5 m of Campanian-Maestrichtian chalk and limestone with only 18.2% recovery, whereas Site 700 recovered 66.8% of a 152.7-m section of Coniacian-Maestrichtian limestone. Preservation of planktonic foraminifers from both sites is moderate in Maestrichtian samples, but worsens with increasing depth in the Campanian. The Northeast Georgia Rise planktonic foraminifers are typical of Late Cretaceous Austral Province faunas described from other southern high-latitude sites; species diversity is low and the assemblages are dominated by species of Heterohelix, Globigerinelloides, Hedbergella, and Archaeoglobigerina. Five species, including Globigerinelloides impensus Sliter, Archaeoglobigerina australis Huber, Archaeoglobigerina mateola Huber, Hedbergella sliteri Huber, and Rugotruncana circumnodifer (Finlay), are considered to be endemic to the Austral Province. Formation of a cool temperate water mass in the circum-Antarctic region, resulting from the final breakup of the Gondwana continents, may have led to increased provincialism of the Austral Province planktonic foraminifers during Campanian-Maestrichtian time. Magnetobiostratigraphic correlation of eight planktonic foraminifer datum events at Hole 700B with ages determined for datums at ODP Leg 113 Holes 689B and 690C (Maud Rise, 65°S) demonstrates regional synchroneity of first and last occurrences within the Austral Province. As was observed at the Maud Rise, several keeled and nonkeeled species previously thought to have been restricted to warmer low-latitude regions first occur later at the Northeast Georgia Rise than at the low-latitude sites. The causes for high-latitude diachroneity among these immigrant species are not clear; neither oxygen and carbon isotope data from the Maud Rise sites nor calcareous nannoplankton distributions for the southern South Atlantic region show conspicuous changes that correlate to the delayed first occurrences.

Analysis of an ancient global warming event, the Paleocene-Eocene thermal maximum, under examination from ODP Holes 113-689B and 113-690B

Spatiotemporal patterns of carbonate dissolution provide a critical constraint on carbon input during an ancient (~55.5 Ma) global warming event known as the Paleocene-Eocene thermal maximum (PETM), yet the magnitude of lysocline shoaling in the Southern Ocean is poorly constrained due to limited spatial coverage in the circum-Antarctic region. This shortcoming is partially addressed by comparing patterns of carbonate sedimentation at the Site 690 PETM reference section to those herein reconstructed for nearby Site 689. Biochemostratigraphic correlation of the two records reveals that the first ~36 ka of the carbon isotope excursion (CIE) signaling PETM conditions is captured by the Site 689 section, while the remainder of the CIE interval and nearly all of the CIE recovery are missing due to a coring gap. A relatively expanded stratigraphy and higher carbonate content at mid-bathyal Site 689 indicate that dissolution was less severe than at Site 690. Thus, the bathymetric transect delimited by these two PETM records indicates that the lysocline shoaled above Site 689 (~1,100 m) while the calcite compensation depth remained below Site 690 (~1,900 m) in the Weddell Sea region. The ensuing recovery of carbonate sedimentation conforms to a bathymetric trend best explained by gradual lysocline deepening as negative feedback mechanisms neutralized ocean acidification. Further, biochemostratigraphic evidence indicates the tail end of the CIE recovery interval at both sites has been truncated by a hiatus most likely related to vigorous production and advection of intermediate waters.

Stratigraphy of the Campanian-Maastrichtian boundary in ODP Hole 122-762C

An integrated framework of magnetostratigraphy, calcareous microfossil bio-events, cyclostratigraphy and d13C stratigraphy is established for the upper Campanian-Maastrichtian of ODP Hole 762C (Exmouth Plateau, Northwestern Australian margin). Bulk-carbonate d13C events and nannofossil bio-events have been recorded and plotted against magnetostratigraphy, and provided absolute ages using the results of the cyclostratigraphic study and the recent astronomical calibration of the Maastrichtian. Thirteen carbon-isotope events and 40 nannofossil bio-events are recognized and calibrated with cyclostratigraphy, as well as 14 previously published foraminifer events, thus constituting a solid basis for large-scale correlations. Results show that this site is characterized by a nearly continuous sedimentation from the upper Campanian to the K-Pg boundary, except for a 500 kyr gap in magnetochron C31n. Correlation of the age-calibrated d13C profile of ODP Hole 762C to the d13C profile of the Tercis les Bains section, Global Stratotype Section and Point of the Campanian-Maastrichtian boundary (CMB), allowed a precise recognition and dating of this stage boundary at 72.15 ± 0.05 Ma. This accounts for a total duration of 6.15 ± 0.05 Ma for the Maastrichtian stage. Correlation of the boundary level with northwest Germany shows that the CMB as defined at the GSSP is ~800 kyr younger than the CMB as defined by Belemnite zonation in the Boreal realm. ODP Hole 762C is the first section to bear at the same time an excellent recovery of sediments throughout the upper Campanian-Maastrichtian, a precise and well-defined magnetostratigraphy, a high-resolution record of carbon isotope events and calcareous plankton biostratigraphy, and a cyclostratigraphic study tied to the La2010a astronomical solution. This section is thus proposed as an excellent reference for the upper Campanian-Maastrichtian in the Indian Ocean.

Neogene Radiolarians in Neogene sediments of the Norwegian Sea

Radiolaria are present in frequencies ranging from rare to abundant and with generally moderate to good preservation quality in Leg 104 sediments younger than 22 Ma. Preservation degrades in progressively younger sediments, and upper Pliocene to mid-Pleistocene radiolaria were found only at Site 644, where sporadic assemblages of moderate to poorly preserved specimens persist to approximately 0.75 Ma. Radiolaria are essentially absent in Leg 104 recovery older than basal Miocene. The stratigraphic ranges of 55 taxa of radiolaria are documented in 451 samples from the biosiliceous recoveries of Holes 642B, 642C, 642D, 643A, and 644A. The stratigraphic ranges of 25 of these species are used as boundary criteria for a new system of 28 Neogene zones and subzones that are used to characterize approximately 72% of the past 22 m.y. of sedimentation on the Vriring Plateau. This new scheme is intended to supercede the NRS zones provisionally proposed in the Leg 104 Initial Reports. The applicability of this regional biozonation beyond the Wring and Iceland Plateaus is not presently known. The radiolaria biostratigraphy serves as a basis for inferring a sequence of hiatuses and faunal overturns that may be associated with sea-level low stands and consequent cold-water isolation of the Norwegian Sea. Twenty-one new taxa are described as follows: Actinomma henningsmoeni, Actinomma livae, Actinomma mirabile, Actinomma plasticum, Ceratocyrtis broeggeri, Ceratocyrtis manumi, Ceratocyrtis stoermeri, Clathrospyris vogti, Corythospyris hispida, Corythospyris jubata sverdrupi, Corythospyris reuschi, Crytocapsella ampullacea, Cyrocapsella kladaros, Gondwanaria japonica kiaeri, Hexalonche esmarki, Larcospira bulbosa, Phormospyris thespios, Pseudodicytophimus amundseni, Spongotrochus vitabilis, Spongurus cauleti, and Tessarastrum thiedei.

Cretaceous planktonic foraminifera of the Kerguelen Plateau

An almost complete Upper Cretaceous sedimentary sequence recently recovered on the Kerguelen Plateau (southern Indian Ocean) during ODP Leg 183 was analysed for planktonic foraminifera in order to refine and integrate the zonal schemes previously proposed for the Southern Ocean area. Detailed biostratigraphic analysis carried out on holes 1135A, 1136A and 1138A (poleward of 50°S palaeolatitude during Late Cretaceous time) has allowed recognition of low and mid-high latitude bioevents, useful for correlation across latitudes, in addition to known Austral bioevents. The low latitude biozonation can be applied to Turonian sediments, because of the occurrence of Helvetoglobotruncana helvetica, which marks the boundary between Whiteinella archaeocretacea and Helvetoglobotruncana helvetica zones. The base of the Whiteinella archeocretacea Zone falls within the uppermost Cenomanian-Turonian black shale level in Hole 1138A. The stratigraphic interval from upper Turonian to uppermost Santonian can be resolved using bioevents recognized in the mid-high latitude sections. They are, in stratigraphic order: the last occurrence of Falsotruncana maslakovae in the Coniacian, the first occurrence of Heterohelix papula at the Coniacian/Santonian boundary, the extinction of the marginotruncanids in the late Santonian, and the first occurrence of Globigerinelloides impensus in the latest (?) Santonian. The remainder of the Late Cretaceous fits rather well in the Austral zonal scheme, except that Globigerinelloides impensus exhibits a stratigraphic range in agreement with its record at the mid-high latitude sections and extends further downwards than previously recorded at southern sites. Therefore, despite the poor recovery in certain intervals and the presence of several hiatuses of local and regional importance as revealed by correlation among holes, a more detailed zonal scheme has been obtained (mainly for the less resolved Turonian-Santonian interval). Remarks on some species often overlooked in literature are also provided.

Isotopic and micropaleontological record of sediments of the Baffin Bay

The oxygen and carbon isotopic compositions of the planktonic foraminifer, Neogloboquadrina pachyderma (sinistral), were determined at 20-cm intervals through the 'composite' top ~ 22 m of sediments at ODP Site 645 (Holes 645B, 645C, 645F, and 645G) and at 10-cm intervals through a 9-m piston core (85-027-016) collected during the Hudson site survey. Quantitative analyses of palynomorphs, notably dinocysts, and of planktonic foraminifers were performed. Organic and nitrogen contents and isotopic composition of nitrogen and carbon in organic matter also were determined. These data provide a high-resolution record of changes that occurred in surface-water masses during the last glacial cycle in Baffin Bay. The basin experienced low planktonic productivity during most of the late Pleistocene, either from dilution in surface water by meltwater discharges from the surrounding ice-sheet or from the presence of a relatively dense sea-ice cover. Peaks of meltwater discharge are indicated by d18O values as low as about 1.5 per mil, correlative d13C- d18O shifts, low concentration of planktonic foraminifers, high concentrations of glacially reworked pre-Quaternary palynomorphs, and low-salinity dinocyst assemblages. As a whole, d18O values ranging between 4.5 and 2.5 per mil allow the establishment of an 18O stratigraphy spanning isotopic stages 5 to 1. Because of the poor core recovery, the general paucity of microflora and microfauna, and the possible occurrence of slumping or debris flow at Site 645, further interpretation remains problematic.

Mineralogy, geochemistry and benthic foraminifera of the Gorrondatxe beach section

A distinctive low-carbonate interval interrupts the continuous limestone-marl alternation of the deep-marine Gorrondatxe section at the early Lutetian (middle Eocene) C21r/C21n Chron transition. The interval is characterized by increased abundance of turbidites and kaolinite, a 3 per mil decline in the bulk d13C record, a >1 per mil decline in benthic foraminiferal d13C followed by a gradual recovery, a distinct deterioration in foraminiferal preservation, high proportions of warm-water planktic foraminifera and opportunistic benthic foraminifera, and reduced trace fossil and benthic foraminiferal diversity, thus recording a significant environmental perturbation. The onset of the perturbation correlates with the C21r-H6 event recently defined in the Atlantic and Pacific oceans, which caused a 2°C warming of the seafloor and increased carbonate dissolution. The perturbation was likely caused by the input of 13C-depleted carbon into the ocean-atmosphere system, thus presenting many of the hallmarks of Paleogene hyperthermal deposits. However, from the available data it is not possible to conclusively state that the event was associated with extreme global warming. Based on our analysis, the perturbation lasted 226 kyr, from 47.44 to 47.214 Ma, and although this duration suggests that the triggering mechanism may have been similar to that of the Paleocene-Eocene Thermal Maximum (PETM), the magnitude of the carbon input and the subsequent environmental perturbation during the early Lutetian event were not as severe as in the PETM.

Mass balance and acceleration of the Greenland ice sheet

Within the last decade, the Greenland ice sheet (GrIS) and its surroundings have experienced record high surface temperatures (Mote, 2007, doi:10.1029/2007GL031976; Box et al., 2010), ice sheet melt extent (Fettweis et al., 2011, doi:10.5194/tc-5-359-2011) and record-low summer sea-ice extent (Nghiem et al., 2007, doi:10.1029/2007GL031138). Using three independent data sets, we derive, for the first time, consistent ice-mass trends and temporal variations within seven major drainage basins from gravity fields from the Gravity Recovery and Climate Experiment (GRACE; Tapley et al., 2004, doi:10.1029/2004GL019920), surface-ice velocities from Inteferometric Synthetic Aperture Radar (InSAR; Rignot and Kanagaratnam, 2006, doi:10.1126/science.1121381) together with output of the regional atmospheric climate modelling (RACMO2/ GR; Ettema et al., 2009, doi:10.1029/2009GL038110), and surface-elevation changes from the Ice, cloud and land elevation satellite (ICESat; Sorensen et al., 2011, doi:10.5194/tc-5-173-2011). We show that changing ice discharge (D), surface melting and subsequent run-off (M/R) and precipitation (P) all contribute, in a complex and regionally variable interplay, to the increasingly negative mass balance of the GrIS observed within the last decade. Interannual variability in P along the northwest and west coasts of the GrIS largely explains the apparent regional mass loss increase during 2002-2010, and obscures increasing M/R and D since the 1990s. In winter 2002/2003 and 2008/2009, accumulation anomalies in the east and southeast temporarily outweighed the losses by M/R and D that prevailed during 2003-2008, and after summer 2010. Overall, for all basins of the GrIS, the decadal variability of anomalies in P, M/R and D between 1958 and 2010 (w.r.t. 1961-1990) was significantly exceeded by the regional trends observed during the GRACE period (2002-2011).

Plio-Pleistocene benthic foraminifera of the Tyrrhenian Sea

Benthic foraminifers from Site 652, Site 653 (Hole 653A), and Site 654 of Leg 107 (Tyrrhenian Sea, Western Mediterranean), which penetrated with more or less good recovery the Plio-Pleistocene stratigraphic interval, were studied in a total of 699 close-spaced samples. A total number of 269 species have been classified and their quantitative distribution in each sample is reported. The benthic foraminifers assemblage is more diversified in Site 654, less diversified in Site 652. Less than a half of the benthic foraminifers species listed from Plio-Pleistocene Italian land sections are present in the coeval deep-sea Tyrrhenian record, in which shallow water species are missing and Nodosarids are poorly represented. A very few species have comparable stratigraphic distribution in the three deep-sea sequences and in Italian land sections when compared against calcareous plankton biostratigraphy. In the same three sites, the first appearance levels of several species are younger and younger, and last appearance levels are earlier and earlier from Site 654 to Site 653 and Site 652. Five biostratigraphic events, biochronologically evaluated and occurring at the same level in the deepsea Tyrrhenian record and in several land sections, have been selected as zonal boundaries of the proposed benthic foraminifers biostratigraphic scheme. The Plio-Pleistocene interval has been subdivided into four biozones and one subzone, recognizable both in the deep-sea and land-based sequences. The Cibicidoides (?) italicus assemblage zone stretches from the base of the Pliocene to the extinction level of the zonal marker, biochronologically evaluated at 2.9 Ma. The Cibicidoides robertsonianus interval zone stretches from the Cibicidoides (?) italicus extinction level to the Pliocene Mediterranean FO of Gyroidinoides altiformis, evaluated at 2.4 Ma. The Gyroidinoides altiformis interval zone stretches from the Mediterranean Pliocene FO of the zonal marker to the appearance level of Articulina tubulosa, evaluated at 1.62 Ma. The Articulina tubulosa assemblage zone stretches from the appearance level of the zonal marker to the Recent. In the Articulina tubulosa biozone, the Hyalinea baltica subzone is proposed. The appearance level of Hyalinea baltica is evaluated at 1.35 Ma, well above the Plio-Pleistocene boundary as defined in the Vrica stratotype section.

Middle Eocene-Early Pliocene Subantarctic planktic foraminiferal biostratigraphy of ODP Site 1090 on the Agulhas Ridge, South Atlantic

The analysis of planktic foraminiferal assemblages from Site 1090 (ODP Leg 177), located in the central part of the Subantarctic Zone south of South Africa, provided a geochronology of a 330-m-thick sequence spanning the Middle Eocene to Early Pliocene. A sequence of discrete bioevents enables the calibration of the Antarctic Paleogene (AP) Zonation with lower latitude biozonal schemes for the Middle-Late Eocene interval. In spite of the poor recovery of planktic foraminiferal assemblages, a correlation with the lower latitude standard planktic foraminiferal zonations has been attempted for the whole surveyed interval. Identified bioevents have been tentatively calibrated to the geomagnetic polarity time scale following the biochronology of Berggren et al. (1995). Besides planktic foraminiferal bioevents, the disappearance of the benthic foraminifera Nuttallides truempyi has been used to approximate the Middle/Late Eocene boundary. A hiatus of at least 11.7 Myr occurs between V78 and V71 m composite depth extending from the Early Miocene to the latest Miocene-Early Pliocene. Middle Eocene assemblages exhibit a temperate affinity, while the loss of several planktic foraminiferal species by late Middle to early Late Eocene time reflects cooling. During the Late Eocene-Oligocene intense dissolution caused impoverishment of planktic foraminiferal assemblages possibly following the emplacement of cold, corrosive bottom waters. Two warming peaks are, however, observed: the late Middle Eocene is marked by the invasion of the warmer water Acarinina spinuloinflata and Hantkenina alabamensis at 40.5 Ma, while the middle Late Eocene experienced the immigration of some globigerinathekids including Globigerinatheka luterbacheri and Globigerinatheka cf. semiinvoluta at 34.3 Ma. A more continuous record is observed for the Early Miocene and the Late Miocene-Early Pliocene where planktic foraminiferal assemblages show a distinct affinity with southern mid- to high-latitude faunas.

Metadata und statistic analysis of archael and bacterial sequences originating from sediments of the Håkon Mosby mud volcano (Z1-Z3, all habitats)

DNA extraction was carried out as described on the MICROBIS project pages (http://icomm.mbl.edu/microbis ) using a commercially available extraction kit. We amplified the hypervariable regions V4-V6 of archaeal and bacterial 16S rRNA genes using PCR and several sets of forward and reverse primers (http://vamps.mbl.edu/resources/primers.php). Massively parallel tag sequencing of the PCR products was carried out on a 454 Life Sciences GS FLX sequencer at Marine Biological Laboratory, Woods Hole, MA, following the same experimental conditions for all samples. Sequence reads were submitted to a rigorous quality control procedure based on mothur v30 (doi:10.1128/AEM.01541-09) including denoising of the flow grams using an algorithm based on PyroNoise (doi:10.1038/nmeth.1361), removal of PCR errors and a chimera check using uchime (doi:10.1093/bioinformatics/btr381). The reads were taxonomically assigned according to the SILVA taxonomy (SSURef v119, 07-2014; doi:10.1093/nar/gks1219) implemented in mothur and clustered at 98% ribosomal RNA gene V4-V6 sequence identity. V4-V6 amplicon sequence abundance tables were standardized to account for unequal sampling effort using 1000 (Archaea) and 2300 (Bacteria) randomly chosen sequences without replacement using mothur and then used to calculate inverse Simpson diversity indices and Chao1 richness (doi:10.2307/4615964). Bray-Curtis dissimilarities (doi:10.2307/1942268) between all samples were calculated and used for 2-dimensional non metric multidimensional scaling (NMDS) ordinations with 20 random starts (doi:10.1007/BF02289694). Stress values below 0.2 indicated that the multidimensional dataset was well represented by the 2D ordination. NMDS ordinations were compared and tested using Procrustes correlation analysis (doi:10.1007/BF02291478). All analyses were carried out with the R statistical environment and the packages vegan (available at: http://cran.r-project.org/package=vegan), labdsv (available at: http://cran.r-project.org/package=labdsv), as well as with custom R scripts. Operational taxonomic units at 98% sequence identity (OTU0.03) that occurred only once in the whole dataset were termed absolute single sequence OTUs (SSOabs; doi:10.1038/ismej.2011.132). OTU0.03 sequences that occurred only once in at least one sample, but may occur more often in other samples were termed relative single sequence OTUs (SSOrel). SSOrel are particularly interesting for community ecology, since they comprise rare organisms that might become abundant when conditions change.16S rRNA amplicons and metagenomic reads have been stored in the sequence read archive under SRA project accession number SRP042162.

Molecular fingerprinting and amplicon sequencing of the bacterial biofilm on settlement tiles deployed along natural pH gradients in Papua New Guinea

Bacterial biofilms provide cues for the settlement of marine invertebrates such as coral larvae, and are therefore important for the resilience and recovery of coral reefs. This study aimed to better understand how ocean acidification may affect the community composition and diversity of bacterial biofilms on surfaces under naturally reduced pH conditions. Settlement tiles were deployed at coral reefs in Papua New Guinea along pH gradients created by two CO2 seeps, and upper and lower tiles surfaces were sampled 5 and 13 months after deployment. Automated Ribosomal Intergenic Spacer Analysis were used to characterize more than 200 separate bacterial communities, complemented by amplicon sequencing of the bacterial 16S rRNA gene of 16 samples. The bacterial biofilm consisted predominantly of Alpha-, Gamma- and Deltaproteobacteria, as well as Cyanobacteria, Flavobacteriia and Cytophaga, whereas putative settlement-inducing taxa only accounted for a small fraction of the community. Bacterial biofilm composition was heterogeneous with approximately 25% shared operational taxonomic units between samples. Among the observed environmental parameters, pH only had a weak effect on community composition (R² ~ 1%) and did not affect community richness and evenness. In contrast, there were strong differences between upper and lower surfaces (contrasting in light exposure and grazing intensity). There also appeared to be a strong interaction between bacterial biofilm composition and the macroscopic components of the tile community. Our results suggest that on mature settlement surfaces in situ, pH does not have a strong impact on the composition of bacterial biofilms. Other abiotic and biotic factors such as light exposure and interactions with other organisms may be more important in shaping bacterial biofilms than changes in seawater pH.